Duplex nested-PCR for detection of small ruminant lentiviruses
| Autor: | Ana Lídia Madeira de Sousa, Sabrina Tainah C. Silva, Gabrielle Rosemblit Martins, Kelma Costa de Souza, Juliana A. Nobre, Rebeca Cavalcante Marinho, Maria Fátima da Silva Teixeira |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
MVV 040301 veterinary sciences viruses lcsh:QR1-502 Sheep Diseases Biology Microbiology Polymerase Chain Reaction DNA sequencing Virus lcsh:Microbiology law.invention 0403 veterinary science 03 medical and health sciences law Small ruminant Animals Multiplex Genetic variability Polymerase chain reaction DNA Primers Goat Diseases Sheep Goats CAEV 04 agricultural and veterinary sciences Virology Standardization 030104 developmental biology Duplex (building) DNA Viral Viruses Lentivirus Infections Nested polymerase chain reaction Research Paper |
| Zdroj: | Brazilian Journal of Microbiology v.49 suppl.1 2018 Brazilian Journal of Microbiology Sociedade Brasileira de Microbiologia (SBM) instacron:SBM Brazilian Journal of Microbiology, Vol 49, Iss suppl 1, Pp 83-92 Brazilian Journal of Microbiology, Volume: 49 Supplement 1, Pages: 83-92, Published: 2018 |
| Popis: | Small ruminant lentiviruses (SRLV) have high genetic variability which results in different viral strains around the world. This create a challenge to design sensible primers for molecular diagnosis in different regions. This work proposes a protocol of duplex nested-PCR for the precise diagnosis of SRLV. The technique was designed and tested with the control strains CAEV Co and MVV 1514. Then, field strains were submitted to the same protocol of duplex nested-PCR. Blood samples of sheep and goats were tested with AGID and nested PCR with specific primers for pol, gag and LTR. The AGID results showed low detection capacity of positive animals, while the nested PCR demonstrated a greater capacity of virus detection. Results demonstrated that LTR-PCR was more efficient in detecting positive sheep samples, whereas gag-PCR allowed a good detection of samples of positive goats and positive sheep. In addition, pol-PCR was more efficient with goat samples than for sheep. Duplex nested PCR performed with standard virus samples and field strains demonstrated that the technique is more efficient for the detection of multiple pro-viral DNA sequences. This study demonstrated a successful duplex nested PCR assay allowing a more accurate diagnosis of SRLV. |
| Databáze: | OpenAIRE |
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